Embryonic Cadaverine Signaling: Implications for Plasma Free Amino Acid and Skeletal Muscle Energy Metabolism in Newly Hatched Chicks.

Cadaverine is a bioactive substance derived from lysine degradation by lysine decarboxylase and has gained attention for its physiological effects. Studies in rodents have revealed its role as a cell growth regulator, particularly intestinal bacterial-produced cadaverine. However, the nutritional and physiological roles of cadaverine during the embryonic period remain unclear, especially considering the immature state of the gut microbiota and digestive functions during this stage. This study explored the potential functions of cadaverine as a nutritional and metabolic signal during chicken embryonic development. Experiments were conducted using an in ovo administration method to evaluate the effects of nutritional bioactive substances on developing chicken embryos. Although there were no observable changes in body or organ weights of newly hatched chicks following in ovo cadaverine administration to day 18 chick embryos, plasma tryptophan, Nτ-methylhistidine, and Nπ-methylhistidine concentrations decreased and the gene expression of insulin/insulin-like growth factor 1 signaling in skeletal muscle was upregulated. These findings imply that cadaverine influences tryptophan metabolism and skeletal muscle catabolism during the embryonic period, suggesting its role as a bioactive factor contributing to energy metabolism signaling in skeletal muscle.

Analysis of prognostic factors in patients with self-expandable metallic stents for treatment of malignant gastric outlet obstruction.

BACKGROUND AND AIM: Self-expandable metallic stents (SEMSs) are widely accepted as a less-invasive treatment for malignant gastric outlet obstruction (GOO). However, the factors related to prognosis and stent dysfunction after SEMS placement are not well known, and we aimed to investigate them. METHODS: This was a single-center retrospective cohort study of 212 malignant strictures in 207 patients. Factors related to prolonged overall survival (OS) and time to recurrent GOO (TRGOO) after SEMS placement were evaluated. RESULTS: Improvement of oral intake was confirmed in 179 patients (86%). The median OS was 65 days. A Cox proportional hazards model revealed that lower cancer stage, lower performance status score at the time of SEMS placement, and administration of chemotherapy after SEMS placement were significant predictive factors for prolonged OS. The median OS was 182 days in the group of SEMS followed by chemotherapy (group A) and 43 days in the group of SEMS alone (group B) (p< .0001). Chemotherapy after SEMS implantation contributed to the prolongation of survival in gastric cancer (hazard ratio (HR), 0.12) and pancreatic cancer (HR, 0.41). Furthermore, the cumulative incidence rates of stent dysfunction on day 120 after SEMS placement were 30% in group A and 61% in group B (p=.03). Notably, the preventive effect of chemotherapy on stent dysfunction was significant in pancreatic cancer. CONCLUSIONS: SEMS is a treatment with high technical and clinical success rate for malignant GOO. Furthermore, subsequent chemotherapy prolongs OS especially in gastric cancer, and TRGOO in pancreatic cancer.

Capacity of Autonomous Sensory Meridian Response on the Reduction of Mental Stress.

In a social environment, various types of stress can be overwhelming. Humans frequently encounter these stressful situations in social life. Stress is divided into physical stress and mental stress; the latter is induced by heavy mental workloads and has become a huge social problem, leading to mental disorders and possibly suicide in the worst scenario. Investigations into monitoring mental stress and reducing stressful conditions are, therefore, important for its prevention. In the present study, we focused on autonomous sensory meridian response (ASMR) sound, which is known to improve the human mental condition through its comforting and relaxing effects. We investigated the effect of ASMR on the mental workload induced by mental tasks by the evaluation of EEG activation patterns in normal subjects. Our results showed a significant decrease in alpha-band activity and a significant increase in gamma (high beta)-band activity under the induction of mental workload by mental tasks compared to the resting condition. When applying ASMR sound, alpha- and gamma-band activity under the induction of mental workload by mental tasks was restored to the level of the resting condition. In conclusion, these results indicate that ASMR sound reduces the mental stress induced by mental workload.

Cryopreservation Competence of Chicken Oocytes as a Model of Endangered Wild Birds: Effects of Storage Time and Temperature on the Ovarian Follicle Survival.

For the conservation of endangered avian species, developing gamete preservation technologies is essential. However, studies in oocytes have not been widely conducted. In this study, assuming that the ovaries are transported to a research facility after death, we investigated the effect of ovary storage on oocytes for the purpose of cryopreserving avian female gametes by using a chicken as a model of endangered avian species. After excision, the ovaries were stored at either a low temperature (4 °C) or room temperature for 1-3 days. Ovarian follicles stored under different conditions for each period were examined by neutral red staining, histology, and gene and protein expression analysis. In addition, the pH of the storage medium after preserving the ovaries was measured. Then, ovarian tissues were vitrified to determine the cryopreservation competence. Storing the ovarian tissues at 4 °C kept the follicles viable and morphologically normal for 3 days with slow decline. In contrast, although different storage temperature did not influence follicle viability and morphology after only 1 day of storage, ovarian tissues stored at room temperature rapidly declined in structurally normal follicles, and viable follicles were rarely seen after 3 days of storage. Gene and protein expression analysis showed that apoptosis had already started on the first day, as shown by the higher expression of CASP9 under room temperature conditions. Furthermore, high expression of SOD1 and a rapid decline of pH in the storage medium under room temperature storage suggested the influence of oxidative stress associated with low pH in this condition on the follicle survivability in hen ovarian tissues. Our cryopreservation study also showed that ovarian tissues stored at 4 °C could recover after cryopreservation even after 3 days of storage. The described storage conditions and cryopreservation methods, which preserve chicken follicle survival, will lay the foundation of ovarian tissue preservation to preserve the fertility of wild female birds.

Metabolism of Imidazole Dipeptides, Taurine, Branched-Chain Amino Acids, and Polyamines of the Breast Muscle Are Affected by Post-Hatch Development in Chickens.

To explore metabolic characteristics during the post-hatch developmental period, metabolomic analyses of breast muscle and plasma were performed in chickens. The most significant growth-related changes in metabolite levels were observed between seven and 28 days of age. Some of these metabolites are essential nutrients or reported as growth-promoting metabolites. In the muscle, two imidazole dipeptides-carnosine and its methylated metabolite, anserine-increased with the development. These dipeptide levels may be, in part, regulated transcriptionally because in the muscle mRNA levels of carnosine synthase and carnosine methylation enzyme increased. In contrast, taurine levels in the muscle decreased. This would be substrate availability-dependent because some upstream metabolites decreased in the muscle or plasma. In branched-chain amino acid metabolism, valine, leucine, and isoleucine decreased in the muscle, while some of their downstream metabolites decreased in the plasma. The polyamines, putrescine and spermidine, decreased in the muscle. Furthermore, mRNA levels associated with insulin/insulin-like growth factor 1 signaling, which play important roles in muscle growth, increased in the muscle. These results indicate that some metabolic pathways would be important to clarify metabolic characteristics and/or growth of breast muscle during the post-hatch developmental period in chickens.

Effect of Stereotaxic Surgery of the Third Ventricle on Growth Performance in Neonatal Chicks.

Feeding behavior and energy metabolism are precisely regulated by humoral and/or neural factors in the central nervous system. In particular, nuclei, such as the arcuate nucleus, ventromedial hypothalamic nucleus, and lateral hypothalamic area located near the third ventricle of the hypothalamus are the centers of feeding and energy metabolism in various vertebrate species, including chickens. In this study, we evaluated the effects of cannulation of the third ventricle on chick growth and feeding behavior in the neonatal stage, to develop a method for local and chronic central nervous system-mediated energy metabolism. Referring to the chick brain atlas, a guide cannula was inserted into the third ventricle of the chick under anesthesia immediately after hatching using a stereotaxic instrument. The chicks that recovered from anesthesia were bred for 11 days under normal feeding management conditions, and then feed intake amount, body weight gain, and metabolic tissue weight were measured. The effects of direct stimulation of the third ventricle with 2-deoxy-D-glucose on the expression level of the immediate-early gene, cFOS, and feed intake in 5-day-old chicks were also evaluated. There were no differences in feed intake, body weight gain, and metabolic tissue weight between 11-day-old cannulated and control chicks. The expression of cFOS mRNA in the ventromedial hypothalamic nucleus was higher than that in the amygdala after the third ventricular administration of 2-deoxy-D-glucose. Additionally, direct third ventricular injection of 2-deoxy-D-glucose attenuated the feeding behavior of chicks for a while. Overall, we speculate that the technique is effective for local and/or chronic stimulation of the nucleus near the third ventricle of the chick hypothalamus, which is important for feed and energy metabolism regulation.

Effects of In Ovo Vitamin D3 Injection on Subsequent Growth of Broilers.

This study was conducted to investigate the influence of in ovo vitamin D3 (Vit D3) administration on growth of broiler chickens when Vit D3 was dissolved in soybean oil. Sixty Ross broiler eggs were incubated at 37.8°C and >60% relative humidity. Distilled water, soybean oil, or Vit D3 (60 IU / 0.5 mL) dissolved in soybean oil, was administered in ovo on Day 18 of incubation. Seven days after hatching, chicks were sexed, and 12 birds (six female and six male) close to the average body weight (BW) of each treatment were selected and their BW continuously recorded until 28 days of age, then sacrificed. Liver and pectoral muscle were collected to determine the mRNA expression of IGF-1 and IGF-1 receptor, and the length of tibia was measured. There were no significant differences in BW, liver weight, or pectoral muscle weight between the groups. However, an interaction was observed between treatments and sexes in the tibia length. In comparison among only males, tibia length in the Vit D3 with oil group was longer than that of the control, but not different from that of the oil group. The same tendency was observed in the hepatic IGF-1 mRNA expression in chicks of either sex, with this effect only being observed after the treatments and not in the control. On the other hand, there was an interaction between treatments and sexes in the mRNA expression of IGF-1 receptor, which was highest in the Vit D3 with oil group in females, but not in males. These results indicated that the in ovo administration of Vit D3 affected IGF-1 receptor mRNA expression without growth.

Usefulness of Urinary Creatinine/Urea Nitrogen Ratio as Indicator of Body Protein Catabolism in Dogs Fed Low Protein Diets.

Low protein diets (LPs) constitute a reportedly effective form of nutritional therapy for canine chronic kidney disease and cirrhosis. These diets have long been feared to result in reduced muscle mass due to protein catabolism. This adverse effect, however, remains largely unrecognized in veterinary medicine as there are no easily applicable catabolism indicators. Therefore, we focused on urinary creatinine, a metabolite of protein in the urine, and examined whether its ratio to urinary urea nitrogen (UCrn/UN) can be used to assess protein catabolism. In Experiment 1, we first consecutively fed seven healthy beagles an LP, standard protein (SP), and high protein (HP) diet for 1 week each and then measured the UCrn/UN ratio at 2-h intervals from fasting to 16 h post-prandially. We consequently found that the UCrn/UN ratio was significantly elevated in the LP pre-prandially and at all post-prandial measurement points (P < 0.01). No significant differences were observed between the SP and HP. Analysis of fasting plasma amino-acid concentrations revealed that the concentration of methionine was significantly lower in the LP than in the other diets (P < 0.05). Although the effects of this change in amino-acid concentration were unclear, the UCrn/UN ratio was considered having increased due to a deficiency in protein and/or amino acids during LP feeding. In Experiment 2, we continuously fed five healthy beagles an LP for 18 weeks and then measured the UCrn/UN ratio as described above. We also measured changes in body composition with computed tomography. At weeks 10 and 18, the fasting UCrn/UN ratio was significantly higher than it was prior to the start of the LP; however, post-prandially, the UCrn/UN ratio decreased to the point that the significant difference disappeared. Muscle mass decreased at weeks 10 and 18. These results suggest that the fasting UCrn/UN ratio could be used as an indicator of protein catabolism in LP feeding. Our experiments thus indicate that examination of potential increases in the UCrn/UN ratio 1 week after introduction of LP feeding to healthy dogs could enable detection of body protein catabolism in long-term feeding of LP before muscle breakdown occurs.

Postural control before and after cerebrospinal fluid shunt surgery in idiopathic normal pressure hydrocephalus.

OBJECTIVES: This study aimed to confirm whether cerebrospinal fluid (CSF) shunting for idiopathic normal-pressure hydrocephalus (iNPH) improves postural instability, and to investigate the relationship between postural control and gait ability. PATIENTS AND METHODS: Twenty-three iNPH patients and 18 age-matched healthy controls (HC) were examined using the timed up and go (TUG) test and a force platform for calculating the center of pressure (COP) trajectory during voluntary multidirectional leaning and quiescent standing. We determined the patients' TUG values and COP trajectories before and after shunt surgery. RESULTS: Postural sway was greater in iNPH patients before shunt surgery and the TUG value was lower in iNPH patients before shunt surgery than in HC. Voluntary COP movements were significantly improved in iNPH patients at 1 week post-surgery, but no significant changes in quiescent standing were found between pre- and post-surgery. Significant correlations were found between the TUG value and voluntary COP movements in iNPH patients before and after surgery, but no significant correlations were observed between the TUG value and quiescent standing. CONCLUSION: Our results indicate that CSF shunting in iNPH patients may improve voluntary postural control and reduce the risk of falling. Impaired voluntary COP control in iNPH patients may reflect the underlying pathophysiological mechanisms of balance disturbance in iNPH.

Estimation of dietary threonine requirement using plasma amino acid concentrations in mature thoroughbreds.

The experiment was conducted to estimate dietary threonine (Thr) requirement using plasma amino acid concentrations as a criterion in mature thoroughbreds. Four adult thoroughbreds were used, and a 4 × 4 Latin square design was used for four dietary Thr levels. Plasma Thr concentration was constant until 0.41%, and then increased rapidly with increasing dietary Thr levels. The Thr requirement was estimated to be 67% of lysine with plasma Thr concentration at four Thr levels.

Selection of regimens for patients with hepatitis C virus genotype/serotype discrepancy.

In the present study, the usefulness of the resistance-associated variant (RAV) analysis to select direct acting antiviral (DAA) drugs for patients with hepatitis C virus (HCV) genotype/serotype discrepancy was evaluated. The core-genotype and serotype were determined in the 559 patients recruited in the study. The RAV analysis and NS5B-genotype determination were performed in the eight patients who exhibited a genotype/serotype discrepancy. One of these patients exhibited a core-genotype 1b/serotype 2, and detection by RAV analysis was possible in this patient. The other seven patients demonstrated a core-genotype 2/serotype 1, and detection using the RAV analysis was possible in four of them. The NS5B-genotype was 1b in all patients in whom detection using the RAV analysis was possible and was other than 1b in patients in whom detection using the RAV analysis was impossible. The RAV analysis could detect RNA sequences specific to genotype 1b in the NS5A region. Therefore, in patients with genotype/serotype discrepancy in whom detection using the RAV analysis is possible, the treatment regimens should be selected based on the assumption that HCV with genome that is highly homologous to genotype 1b is present in the NS5A region.

Effects of Dietary Methionine or Arginine Levels on the Urinary Creatinine Excretion in Broiler Chicks.

Two experiments were conducted to evaluate the usefulness of urinary creatinine levels as a criterion for the estimation of protein and amino acid requirements in poultry. Here we studied the effects of dietary precursor levels of creatinine, methionine and arginine, on urinary creatinine excretion in experiments. Both experiments used 15 Chunky broilers chicks that were 8 days old. The chicks were assigned to three dietary groups, with five chicks each, and were fed an experimental diet for 7 days. The experimental diets mainly consisted of corn and soybean meal, and contained deficient, adequate, or excessive methionine and arginine levels in experiments 1 and 2, respectively. Excreta were collected for the last 3 days of the feeding trial, and chicks were terminated by dislocation of the neck at the end of the feeding trial to collect their livers. Creatinine concentration in the excreta and hepatic l-arginine-glycine amidinotransferase (AGAT) activities were determined. Urinary creatinine levels increased with increasing both dietary methionine and arginine levels from deficient to adequate recommended by Japanese feeding standard (P<0.05), and then remained constant in experiments 1 and 2, respectively. The hepatic AGAT activity decreased when both dietary creatinine precursors levels were increased from deficient to adequate levels (p<0.05), and then remained constant. These results suggested that creatinine excretion was changed with both increasing dietary methionine and arginine, dose-dependently.

[The attempts and current status of cancer rehabilitation at Osaka Medical College Hospital].

We introduced an attempt at cancer rehabilitation at Osaka Medical College Hospital. We also reported trends in the clinical department that ordered the cancer rehabilitation, and the days needed to consult the rehabilitation department after hospitalization for 1,028 patients who needed rehabilitation from January to June 2012. The number of rehabilitation orders for cancer patients has increased in comparison with the same period during 2009, and the percentage of cancer rehabilitation orders has also increased, both in total and in each clinical department consulted. In addition, clinical departments that introduced a rehabilitation schedule along with their treatments ordered cancer rehabilitations much earlier than those departments without such a schedule. In future, to start cancer rehabilitation at an earlier stage, we should endeavor to create awareness of the importance of cancer rehabilitation and the introduction of a rehabilitation schedule along with cancer treatments.

[A case of epithelioid hemangioendothelioma of the liver: detection by FDG-PET].

A 57 year-old woman was admitted for focal accumulation of 18F-fluorodeoxyglucose (FDG) in the liver detected by positron emission tomography (PET). A 25- mm hypovascular tumor was detected by computed tomography. Tumor biopsy revealed many atypical cells with positive staining for factor VIII-related-antigen in sinusoids. Right lobectomy was performed and the tumor was diagnosed as epithelioid hemangioendothelioma (EHE) pathologically. We demonstrated that FDG-PET was useful for the diagnosis of EHE and making deciding on therapeutic strategy.

Expression of mRNA for 3HADH in manipulated embryos to produce germline chimeric chickens.

Germline chimeric chickens were produced by the transfer of primordial germ cells (PGCs) or blastoderm cells. The hatchability of eggs produced by transfer of exogenous PGCs is usually low. The purpose of the present study was investigated to express (3-hydroxyacyl CoA dehydrogenase) 3HADH which is a limiting enzyme in the beta-oxidation of fatty acids for hatching energy. Manipulations of both donor and recipient eggshells were as follows. A window approximately 10 mm in diameter was opened at the pointed end of the eggs at stage 12-15 days incubation. Donor PGCs, taken from the blood vessels of donor embryos from fertilized eggs at the same stage of development, were injected into the blood vessels of recipient embryos. The muscles of chicks in the eggs with transferred PGCs were removed after 20 days of incubation. A cDNA was prepared from the total RNA. The expression of 3HADH in the manipulated embryos was investigated using real-time PCR analysis. Real-time PCR analysis showed that expression of 3HADH was reduced in the muscles of manipulated embryos.

Effect of nociceptin/orphanin FQ on feeding behavior and hypothalamic neuropeptide expression in layer-type chicks.

Nociceptin/orphanin FQ (N/OFQ) was identified in 1995 as the endogenous ligand for the orphan G(i)/G(o)-coupled opioid receptor-like 1 receptor (NOP(1)). Exogenous N/OFQ increases food intake in mammals, but its effect and mode of action in chicks are not fully known. We report herein that N/OFQ (5.0 nmol) has a stimulatory effect on food intake in layer-type chicks over a 2-h period after intracerebroventricular (icv) injection. Thirty minutes after central injection of N/OFQ (5.0 nmol) the concentration of agouti-related protein (AGRP) mRNA in the diencephalon increased, while cocaine- and amphetamine-regulated transcript (CART) mRNA decreased. However, concentrations of neuropeptide Y, proopiomelanocortin and glutamate decarboxylase mRNAs, and of catecholamines and excitatory amino acids were not affected. Simultaneous administration of alpha-melanocyte stimulating hormone (alpha-MSH: 1.0 pmol), a competitor of AGRP, completely blocked the orexigenic effect of N/OFQ (5.0 nmol). These data suggest that N/OFQ functions in layer chicks as an orexigenic peptide in the central nervous system, and that the AGRP and the CART neurons may mediate this function, as in mammals.

Removal of volatile fatty acids with immobilized Rhodococcus sp. B261.

The removal of aqueous volatile fatty acids (VFA) in wastewater and spoiled waste-foods by immobilized Rhodococcus sp. B261 was investigated. The n-valeric acid (0.5%) was completely removed within 25 h under the following conditions; solution pH, 8.0; air flow rate, 0.2 l/min; superficial velocity, 0.96 h(-1); temperature, 37 degrees C. Under the optimized conditions, the acetic (8525 ppm), propionic (7310 ppm) and n-butyric (4360 ppm) except n-valeric (2572 ppm) acids from the wastewater were completely removed by immobilized Rhodococcus sp. B261 in 24 h. The acetic (7810 ppm), propionic (8942 ppm) and butyric (5730 ppm) acids from the solution of spoiled waste-foods were effectively removed by immobilized Rhodococcus sp. B261 from 48 h within 60 h but n-valeric acid (3625 ppm) took 72 h.

Purification and characterization of inulinase from Aspergillus niger AF10 expressed in Pichia pastoris.

The inuA1 gene encoding an exoinulinase from Aspergillus niger AF10 was expressed in Pichia pastoris, and the recombinant enzyme activity was 316U/ml in a 5L fermentor, with the inulinase protein accounting for 35% of the total protein of fermentation broth. The hydrolysis rate of inulin can reach 92%, with a 25U/g inulin enzyme addition, and 90% of fructose content after 6h. Glucose can significantly inhibit the enzymatic hydrolysis of inulin. This is the first report of glucose inhibition of inulinase-catalyzed hydrolysis.

Adenoid cystic carcinoma associated with salivary duct cyst in the sublingual gland.

We described an extremely rare case of adenoid cystic carcinoma associated with salivary duct cyst in the sublingual gland of a 40-year-old Japanese woman. The tumor was growing from the cyst wall and almost occluded the cyst lumen. The epithelium lining the cyst lumen contained both keratin 19-positive cells and alpha-smooth muscle actin-positive cells, indicating the cyst being derived from the acinus/intercalated duct of the sublingual gland. Therefore, our case has presented for the first time a direct evidence that adenoid cystic carcinoma arises from acinus/intercalated duct.